Observation of superoxide production during catalysis of Bacillus subtilis oxalate decarboxylase at pH 4 |
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| Affiliation: | 1. Department of Applied Sciences, Chandigarh Engineering College, Landran, Mohali-140307, India;2. Department of Physics, Chaudhary Devi Lal University, Sirsa-125055, India;3. Department of Mathematics, National Institute of Technology, Hamirpur-177005, India;1. Hospital Universitario de Canarias, Servicio de Cardiología, Santa Cruz de Tenerife, Spain;2. Facultad de Ciencias de la Salud, Universidad Europea de Canarias, La Orotava, Santa Cruz de Tenerife, Spain;3. Centro de Investigación Atmosférica de Izaña (CIAI), AEMET. Unidad Asociada al CSIC, Santa Cruz de Tenerife. Spain;4. Departamento de Ciencias Medicas Básicas (Unidad de Fisiología), Universidad de La Laguna, Santa Cruz de Tenerife, Spain;5. Hospital Universitario Central de Asturias, Área del Corazón, Oviedo, Spain |
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| Abstract: | This contribution describes the trapping of the hydroperoxyl radical at a pH of 4 during turnover of wild-type oxalate decarboxylase and its T165V mutant using the spin-trap BMPO. Radicals were detected and identified by a combination of EPR and mass spectrometry. Superoxide, or its conjugate acid, the hydroperoxyl radical, is expected as an intermediate in the decarboxylation and oxidation reactions of the oxalate monoanion, both of which are promoted by oxalate decarboxylase. Another intermediate, the carbon dioxide radical anion was also observed. The quantitative yields of superoxide trapping are similar in the wild type and the mutant while it is significantly different for the trapping of the carbon dioxide radical anion. This suggests that the two radicals are released from different sites of the protein. |
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| Keywords: | Oxalate decarboxylase Spin trapping BMPO Superoxide Carbon dioxide radical |
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